Linearize pYPKpw with EcoRV resulting in the linearized vector.
Perform the following 2 PCR reactions:
primers 577, 778 and pYPK0_RPL12Btp_CiGXF1_TDH3tp => 3246bp_PCR_prod
primers 775, 578 and pYPK0_TDH3tp_PsXYL2_PGI1tp => 3196bp_PCR_prod
Mix the DNA fragments and transform a S. cerevisiae ura3 mutant. The DNA fragments will be assembled by in-vivo homologous recombination:
-|pYPKpw|124 | \/ | /\ | 124|3246bp_PCR_prod|712 | \/ | /\ | 712|3196bp_PCR_prod|242 | \/ | /\ | 242- | | ----------------------------------------------------